The effect of SUPT4H knockdown on allelic HTT expression using human neurons derived from Huntington's disease iPSCs
| Autor: | Ya-Hsien Tseng, 曾雅嫺 |
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| Rok vydání: | 2017 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 105 Huntington’s disease (HD) is caused by abnormal expansion of CAG tri-nucleotide repeats in the first exon of Huntingtin (HTT) gene, which produces gene products with detrimental effects on neurons. HD, a dominantly progressive neurodegenerative disorder, is clinically associated with movement, cognitive, and emotional impairments. The pathological hallmark of disease is the atrophy of striatum, which is consist of more than 95% GABAergic medium spiny neurons (MSNs). The transcription elongation factor, Spt4, is involved in the process of genes transcription by enhancement of RNA polymerase II processivity on DNA templates. Our previous study, using yeast as a model system, showed that SPT4 inactivation results in a preferential transcription reduction of genes containing a long stretch of CAG tri-nucleotide repeats, while it only marginally affects the expression of genes with short or no CAG repeats. In order to investigate the effect of SUPT4H, the ortholog of yeast SPT4, on the expression of HTT alleles in HD, we introduced a shRNA specifically against SUPT4H under the control of Tet-on system in induced pluripotent stem cells (iPSCs) that are derived from HD patients. Using the engineered iPSCs and GABAergic neurons differentiated from the iPSCs, we demonstrated that the expression of mutant HTT is reduced upon the knockdown of SUPT4H. More importantly, in HD-iPSCs and their differentiated GABAergic neurons, mutant HTT is affected in a greater extent than its wild-type counterpart. These results suggest that preferential reduction of mutant HTT in GABAergic neurons of HD patients is achievable through SUPT4H knockdown. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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